乙烯利暴露对早孕小鼠子宫内膜蜕膜化的影响

Effects of Ethephon Exposure on Endometrial Decidualization in Mice during Early Pregnancy

该研究主要探讨乙烯利(ethephon,ETH)暴露对早孕小鼠子宫内膜蜕膜化的影响。从孕第一天开始每天经口灌胃给予CD1小鼠0、71.25、142.5和285 mg/kg ETH后,于孕第七天处死小鼠。观察子宫胚胎着床数量,记录子宫重量及体重,采用酶联免疫吸附实验(enzyme linked immunesorbent assay,ELISA)检测孕鼠血清雌孕激素水平,RT-PCR、Western blot和免疫组化法(immunohistochemistry,IHC)检测HOXA10、COX2和BMP2等蜕膜化标记分子的mRNA和蛋白表达水平。构建假孕小鼠体内人工诱导蜕膜化模型,RT-PCR检测蜕膜化标记分子HOXA10、COX2和BMP2的mRNA表达水平。结果表明,在285 mg/kg ETH暴露下,小鼠孕第七天子宫胚胎着床数量显著降低(P<0.001),其绝对子宫重量和相对子宫重量均显著低于对照组(P<0.001)。RT-PCR结果显示,285 mg/kg ETH暴露组BMP2和HOXA10 mRNA表达水平显著低于对照组(P<0.001),COX2表达水平显著高于对照组(P<0.001)。Western blot结果显示,与对照组相比,285 mg/kg ETH暴露组其子宫内膜蜕膜化标记分子HOXA10蛋白表达水平显著降低,COX2、MMP9、PR表达水平显著升高。IHC结果显示,与对照组相比,285 mg/kg ETH暴露组其子宫内膜蜕膜化标记分子HOXA10和BMP2蛋白表达水平显著降低。ELISA检测结果表明,285 mg/kg ETH暴露组其血清孕激素水平显著降低(P<0.001)。体内人工诱导蜕膜化模型检测结果显示,ETH暴露组人工诱导蜕膜化反应程度降低,诱导侧子宫重量与非诱导侧子宫重量之比显著低于对照组,蜕膜化标记分子HOXA10和BMP2 mRNA表达水平显著降低。该研究结果表明,孕早期ETH暴露会损害小鼠子宫内膜蜕膜化。

This study focused on whether ETH exposure had adverse effects on endometriosis decidualization in early pregnancy.In this study,on GD1(gestational day 1),CD1 mice were given 0,71.25,142.5,and 285 mg/kg of ETH by continuous oral feeding.On GD7,mice were killed.Uterine tissue was collected and the numbers of implanted embryo were observed.Serum was collected for estrogen and progesterone detection by ELISA.mRNA expression levels of decidual markers such as HOXA10,BMP2 and COX2 were detected by RT-PCR.Protein expression levels of HOXA10,BMP2 and COX2 were detected by immunohistochemistry and Western blot.The induced decidualization model of mice in vivo was established.mRNA expression levels of decidual markers were detected by RT-PCR.Under 285 mg/kg ETH exposure,the numbers of implanted embryos on GD7 were significantly decreased and the absolute and relative uterine weights on GD7 were significantly reduced.RT-PCR results showed BMP2 and HOXA10 mRNA expression were significantly reduced while COX2 mRNA expression was significantly increased compared to control group in the 285 mg/kg ETH exposed group.Western blot result showed that compared with control group,the decidual markers of HOXA10 protein expression was significantly down-regulated while COX2,MMP9 and PR were significantly up-regulated under 285 mg/kg ETH exposed group.Immunohistochemistry result showed HOXA10 and BMP2 protein expression was significantly decreased under 285 mg/kg ETH exposed group.Artifical decidualization was detected in the uteri of the control,but weakened decidualization response was observed in the ETH-treated group.The decidual markers of HOXA10 and BMP2 mRNA were significantly decreased.Serum progesterone levels also decreased obviously by ELISA.Suggested exposure to ETH in early pregnancy had adverse effects on endometrial decidualization in mice.